Functional nucleic acid molecule and use thereof

Abstract

There is disclosed a trans-acting functional nucleic acid molecule comprising a eukaryotic target binding sequence comprising a sequence reverse complementary to a target mRNA sequence for which protein translation is to be enhanced, and a regulatory sequence comprising an internal ribosome entry site (IRES) sequence or an internal ribosome entry site (IRES) derived sequence and enhancing translation of the target mRNA sequence, wherein the regulatory sequence is located 3′ of the target binding sequence.

The invention claimed is: 
     
       1. A trans-acting functional nucleic acid molecule comprising:
 a target binding sequence comprising a sequence reverse complementary to a eukaryotic target mRNA sequence for which protein translation is to be enhanced; and 
 a regulatory sequence comprising an internal ribosome entry site (IRES) sequence or an internal ribosome entry site (IRES) derived sequence and enhancing translation of the target mRNA sequence, 
 
       wherein the regulatory sequence is located 3′ of the target binding sequence. 
     
     
       2. The trans-acting functional nucleic acid molecule according to  claim 1 , wherein the target binding sequence consists, from 3′ to 5′, of a sequence reverse complementary to 1 to 50 nucleotides of the 5′ untranslated region (5′ UTR) and 1 to 40 nucleotides of the coding sequence (CDS) of the target mRNA sequence. 
     
     
       3. The trans-acting functional nucleic acid molecule according to  claim 2 , wherein the target binding sequence consists, from 3′ to 5′, of a sequence reverse complementary to 10 to 45 nucleotides of the 5′ untranslated region (5′ UTR) and 2 to 6 nucleotides of the coding sequence (CDS) of the target mRNA sequence. 
     
     
       4. The trans-acting functional nucleic acid molecule according to  claim 1 , wherein the IRES sequence or IRES derived sequence is oriented, in the trans-acting functional nucleic acid molecule, in direct orientation relative to the 5′ to 3′ orientation of the functional nucleic acid molecule. 
     
     
       5. The trans-acting functional nucleic acid molecule according to  claim 1 , wherein the IRES sequence or IRES derived sequence is a sequence with 75% homology to a sequence selected from the group consisting of SEQ ID NO:36 to SEQ ID NO:65. 
     
     
       6. The trans-acting functional nucleic acid molecule according to  claim 5 , wherein the IRES sequence or IRES derived sequence is a sequence with 90% homology to a sequence selected from the group consisting of SEQ ID NO:36 to SEQ ID NO:65. 
     
     
       7. The trans-acting functional nucleic acid molecule according to  claim 6 , wherein the IRES sequence or IRES derived sequence is a sequence selected from the group consisting of SEQ ID NO:36 to SEQ ID NO:65. 
     
     
       8. The trans-acting functional nucleic acid molecule according to  claim 1 , wherein the trans-acting functional nucleic acid molecule is an RNA molecule or a modified RNA molecule. 
     
     
       9. The trans-acting functional nucleic acid molecule according to  claim 1 , further comprising a spacer sequence between the target binding sequence and the regulatory sequence. 
     
     
       10. A DNA molecule encoding the trans-acting functional nucleic acid molecule according to  claim 1 . 
     
     
       11. An expression vector comprising the DNA molecule according to  claim 10 . 
     
     
       12. A method for enhancing protein translation, the method comprising transfecting into a cell (a) the trans-acting functional nucleic acid molecule according to  claim 1 , (b) a DNA molecule encoding the trans-acting functional nucleic acid molecule according to  claim 1 , or (c) an expression vector comprising a DNA molecule encoding the trans-acting functional nucleic acid molecule according to  claim 1 . 
     
     
       13. A composition comprising (a) the trans-acting functional nucleic acid molecule according to  claim 1 , (b) a DNA molecule encoding the trans-acting functional nucleic acid molecule according to  claim 1 , or (c) an expression vector comprising a DNA molecule encoding the trans-acting functional nucleic acid molecule according to  claim 1 . 
     
     
       14. A method for enhancing translation of a target mRNA sequence, the method comprising hybridizing the target mRNA sequence to (a) the trans-acting functional nucleic acid molecule according to  claim 1 , (b) a DNA molecule encoding the trans-acting functional nucleic acid molecule according to  claim 1 , or (c) an expression vector comprising a DNA molecule encoding the trans-acting functional nucleic acid molecule according to  claim 1 . 
     
     
       15. A method for treating a genetic disease caused by down-regulation of a protein-coding mRNA, the method comprising administering (a) the trans-acting functional nucleic acid molecule according to  claim 1 , (b) a DNA molecule encoding the trans-acting functional nucleic acid molecule according to  claim 1 , or (c) an expression vector comprising a DNA molecule encoding the trans-acting functional nucleic acid molecule according to  claim 1 . 
     
     
       16. A method for treating a genetic or sporadic disease where reduced gene dosage is detrimental, the method comprising administering (a) the trans-acting functional nucleic acid molecule according to  claim 1 , (b) a DNA molecule encoding the trans-acting functional nucleic acid molecule according to  claim 1 , or (c) an expression vector comprising a DNA molecule encoding the trans-acting functional nucleic acid molecule according to  claim 1 .